Computational design and engineering of an Escherichia coli strain producing the nonstandard amino acid para-aminophenylalanine

Zomorrodi, Ali R.; Hemez, Colin; Arranz-Gibert, Pol; Wu, Terrence; Isaacs, Farren J.; Segrè, Daniel

doi:10.1016/j.isci.2022.104562

Title: Computational design and engineering of an Escherichia coli strain producing the nonstandard amino acid para-aminophenylalanine

Journal Article · Thu Jun 09 00:00:00 EDT 2022 · iScience

DOI:https://doi.org/10.1016/j.isci.2022.104562· OSTI ID:1889425

Zomorrodi, Ali R. ^[1]; Hemez, Colin ^[2]; Arranz-Gibert, Pol ^[2]; Wu, Terrence ^[3]; Isaacs, Farren J. ^[2];

^[4]

Massachusetts General Hospital, Boston, MA (United States); Harvard Medical School, Boston, MA (United States); Boston Univ., MA (United States)
Yale Univ., New Haven, CT (United States); Yale Univ., West Haven, CT (United States)
Yale Univ., West Haven, CT (United States)
Boston Univ., MA (United States)

Introducing heterologous pathways into host cells constitutes a promising strategy for synthesizing nonstandard amino acids (nsAAs) to enable the production of proteins with expanded chemistries. However, this strategy has proven challenging, as the expression of heterologous pathways can disrupt cellular homeostasis of the host cell. Here, we sought to optimize the heterologous production of the nsAA para-aminophenylalanine (pAF) in Escherichia coli. First, we incorporated a heterologous pAF biosynthesis pathway into a genome-scale model of E. coli metabolism and computationally identified metabolic interventions in the host’s native metabolism to improve pAF production. Next, we explored different approaches of imposing these flux interventions experimentally and found that the upregulation of flux in the chorismate biosynthesis pathway through the elimination of feedback inhibition mechanisms could significantly raise pAF titers (~20-fold) while maintaining a reasonable pAF production-growth rate trade-off. Overall, this study provides a promising strategy for the biosynthesis of nsAAs in engineered cells.

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Research Organization:: Lawrence Berkeley National Lab. (LBNL), Berkeley, CA (United States)

Sponsoring Organization:: USDOE Office of Science (SC), Biological and Environmental Research (BER); Defense Advanced Research Projects Agency (DARPA); Netherlands Organization for Scientific Research (NWO); National Science Foundation (NSF)

Grant/Contract Number:: AC02-05CH11231; HR0011-15-C-0091; EF-1935120

OSTI ID:: 1889425

Journal Information:: iScience, Vol. 25, Issue 7; ISSN 2589-0042

Publisher:: ElsevierCopyright Statement

Country of Publication:: United States

Language:: English

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Title: Computational design and engineering of an Escherichia coli strain producing the nonstandard amino acid para-aminophenylalanine

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